Saturday, March 9, 2013

Indexing Samples

Here is my earlier plan for multiplexing the samples:
Lane 1:                      Lane 2:
BBM1+2+3              BBM3+4+5
BBF1+2+3               BBF3+4+5
BSM1+2+3              BSM3+4+5
BSF1+2+3               BSF3+4+5
SBM1+2+3              SBM3+4+5
SBF1+2+3               SBF3+4+5
SSM1+2+3              SSM3+4+5
SSF1+2+3               SSF3+4+5

Each of these numbers will have it's own unique barcode. But as I only have 24 barcodes, some will have to be repeated between the lanes. For example, BBM3 must go in both lanes and will have the same barcode. This will force BBM1 and BBM4 to share a barcode, same with BBM2 and BBM5. It will end up with barcodes 1-16 in each lane but on different samples, and barcodes 17-24 in each lane and on the same sample.

Here is a schematic of my plan:
    
Sample ID
Illumina Barcode Number
 Reference Number (above)
Lane
1
BB.BB15.3M 
1
BBM1
1
2
BB.SS70.3M
2
BSM1
1
3
SS.BB20.2M
3
SBM1
1
4
SS.SS82.1M
4
SSM1
1
5
BB.BB15.4F
5
BBF1
1
6
BB.SS70.2F
6
BSF1
1
7
SS.BB20.6F
7
SBF1
1
8
SS.SS82.2F
8
SSF1
1
9
BB.BB77.2M
9
BBM2
1
10
BB.SS70.5M
10
BSM2
1
11
SS.BB25.5M
11
SBM2
1
12
SS.SS87.1M
12
SSM2
1
13
BB.BB77.1F
13
BBF2
1
14
BB.SS70.4F
14
BSF2
1
15
SS.BB25.3F
15
SBF2
1
16
SS.SS87.3F
16
SSF2
1
17
BB.BB86.2M
17
BBM3
both
18
BB.SS72.2M
18
BSM3
both
19
SS.BB25.3M
19
SBM3
both
20
SS.SS88.1M
20
SSM3
both
21
BB.BB86.1F
21
BBF3
both
22
BB.SS72.1F
22
BSF3
both
23
SS.BB29.1F
23
SBF3
both
24
SS.SS88.3F
24
SSF3
both
25
BB.BB87.6M
1
BBM4
2
26
BB.SS73.3M
2
BSM4
2
27
SS.BB20.8M
3
SBM4
2
28
SS.SS89.2M
4
SSM4
2
29
BB.BB87.1F
5
BBF4
2
30
BB.SS73.1F
6
BSF4
2
31
SS.BB24.2F
7
SBF4
2
32
SS.SS89.1F
8
SSF4
2
33
BB.BB77.3M
9
BBM5
2
34
BB.SS71.3M
10
BSM5
2
35
SS.BB22.4M
11
SBM5
2
36
SS.SS91.1M
12
SSM5
2
37
BB.BB90.1F
13
BBF5
2
38
BB.SS73.2F
14
BSF5
2
39
SS.BB24.3F
15
SBF5
2
40
SS.SS91.4F
16
SSF5
2

Furthermore, I will need to prepare these in even batches to avoid any weird batch effects. I will therefore do them in 5 batches of 8. Each batch will have one male and one female from each cross type.

I'll start the first batch (numbers 1-8) tomorrow.


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